This study details a rat fixation method suitable for photographing acupuncture and infrared thermography. Furthermore, it demonstrates that electroacupuncture at sensitized acupoints inhibits local inflammatory pain.
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Method Article
* These authors contributed equally
This study details a rat fixation method suitable for photographing acupuncture and infrared thermography. Furthermore, it demonstrates that electroacupuncture at sensitized acupoints inhibits local inflammatory pain.
Knee osteoarthritis (KOA), a leading cause of musculoskeletal pain and functional disability worldwide, remains challenging to manage due to the limited efficacy of current pharmacological and surgical interventions in achieving sustained pain relief. Although acupuncture has demonstrated clinical promise in alleviating KOA-related pain, its underlying mechanisms require further elucidation. Application of acupuncture to sensitized acupoints can markedly enhance the effectiveness of therapy. This protocol establishes a rat model of KOA through intra-articular injection of sodium monoiodoacetate (MIA) and systematically describes an acupuncture protocol targeting sensitized acupoints, including restraint methods, acupoint localization, and acupuncture parameters. Therapeutic efficacy was comprehensively evaluated through pain behavioral assessments, infrared thermographic analysis of the knee joint, inflammatory cytokine detection in serum, and histopathological examination of cartilage tissue. Results indicate that acupuncture at sensitized acupoints significantly alleviates pain phenotypes in MIA-induced model rats by inhibiting the release of pro-inflammatory mediators, blocking nociceptive signal transmission, improving blood microcirculation, regulating local joint temperature, and delaying joint degeneration.
Knee osteoarthritis (KOA), a prevalent degenerative joint disorder, is characterized by chronic inflammation and whole-joint pathology, including cartilage erosion, subchondral bone remodeling, osteophyte development, and synovial inflammation1,2. Clinically, patients typically present with pain, muscle weakness, joint instability, transient morning stiffness, and progressive functional limitations. KOA imposes a substantial global disease burden, affecting approximately 365 million individuals with functional disability, with epidemiological models predicting a 74.9% escalation in case numbers from 2020 basel....
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All experimental protocols were approved and rigorously supervised by the Animal Ethics Committee of Beijing University of Chinese Medicine (ID: bucm-2024091102-3197), and all procedures strictly adhered to the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals. Thirty male Sprague-Dawley rats (weight range: 210-250 g) were used in this study. The reagents and equipment used are listed in the Table of Materials.
1. Experimental animals and groups
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No animals died during the entire experiment. Results of the operation procedure duration and needle detachment count for each rat in both the NF and EA groups indicated that the EA group, following restriction with nylon cable ties, experienced reduced operation time and fewer needle detachments (P < 0.001; Figure 1H,I). Analysis of acupoint pain thresholds revealed no statistically significant differences in mechanical pain sensitivity between GB34, ST36, and n.......
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MIA, a metabolic inhibitor, selectively disrupts cellular aerobic glycolysis, thereby inducing chondrocyte apoptosis17,18,19. Intra-articular MIA administration results in chondrocyte depletion and subsequent articular cartilage degeneration, characterized by morphological irregularities and histopathological alterations that closely resemble human OA pathology20. Thus, it is widely utilized in research o.......
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All authors have declared no potential conflicts of interest.
This research was supported by the National Key R&D Program of China (grant 2023YFC3502700) and the Fundamental Research Funds for the Central Universities (2025-JYB-XJSJJ014)
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| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| 0-10ul Adjustable-Volume Pipettor | Servicebio | IC012395160823 | |
| 0-200ul Adjustable-Volume Pipettor | Servicebio | IC021029160323 | |
| 1.5 ml eppendorf tubes | Servicebio | EP-150-M | |
| 10ul micropipette tip | Servicebio | TP-10 | |
| 1ml syringe | Shanghai Zhiyu Medical Equipment Co., LTD | ZSQ-1 | |
| 200ul micropipette tip | Servicebio | TP-200 | |
| 22-gauge needle | Kruuse | 112416c | |
| 2ml eppendorf tubes | Servicebio | EP-150-M | |
| 5ml red blood tube | Shanghai Xinle Biotechnology Co. LTD | CXG-5R | |
| 75% alcohol | Shandong Anjie Gaoke Disinfection Technology Co. LTD | 75-500ml | |
| Adherent slide | CITOTEST | 188105 | |
| Automatic Plate Washer | Rayto | RT-3100C | |
| Blood collection needle | Jiangxi Ruibang Industrial Group Co. LTD | CXZ-07 | |
| Bone tissue safranin and solid green staining solution | Shanghai Yuan Ye Biotechnology Co., Ltd. | R32730 | |
| Cover glass | Citotest Labware Manufacturing Co.,Ltd | 10212432C | |
| Dehydrator | DIAPATH | Donatello | |
| Electronic balance | Mettler-Toledo | ME203E/02 | |
| Electrothermal blast drying oven | Labotery | GEL-70 | |
| Embedding machine | Wuhan Junjie Electronics Co., Ltd | JB-P5 | |
| Environmental Friendly Dewaxing Transparent Liquid | Servicebio | G1128-1L | |
| Enzyme label detector | BioTeK | Epoch | |
| Ethanol | SCRC | 100092683 | |
| Ethylenediamine tetraacetic acid | Macklin | 60-00-4 | |
| Fat-free cotton ball | Caoxian Hualu Sanitary Material Co. LTD | TZMQ-500 | |
| High speed refrigerated centrifuge | Dragon | D3024R | |
| High Speed Tissue Grinder | Servicebio | KZ-III-F | |
| Hot Plate Analgesiometer | Kew basis | KW-CT-1 | |
| Imaging system | Nikon | NIKON DS-U3 | |
| Infrared thermal imager | Testo AG | Testo 865 | |
| Iodophor | Shandong Anjie Gaoke Disinfection Technology Co. LTD | DF-500 | |
| IR Soft | Testo AG | ||
| Isoflurane | RWD life science | R510-22-10 | |
| Latex gloves | Beijing Ruijing Latex Products Co. LTD | RJJC-S | |
| Mechanical Paw Poking Test Device | Kew basis | KW-RB | |
| MIA | Sigma | 206-165-7 | |
| Neutral gum | SCRC | 10004160 | |
| Normal butanol | SCRC | 100052190 | |
| Normal saline solution | SHIMEN | SLYS-500ml | |
| Paraffin liquid | Macklin | 8012-95-1 | |
| Paraformaldehyde Fixative (Neutral) | Servicebio | G1101 | |
| Pathology slicer | Shanghai Leica Instrument Co., Ltd | RM2016 | |
| Rat IL-1 beta ELISA Kit GER0002-96t | Servicebio | AC30253754 | |
| Rat TNF-alpha ELISA Kit GER0004-96t | Servicebio | AC40253711 | |
| SD rats | Beijing Sibefei Biotechnology Co., Ltd | SCXK (jing) 2024-0001 | |
| Small Animal Anesthesia Machine | YuYAN | ABM | |
| Surgical mask | winner | WJKZ-1 | |
| Tissue spreader | Zhejiang Kehua Instrument Co., Ltd | KD-P | |
| Ultrapure Water Polishing System | Aiken water El | AK-RO-C2 | |
| Upright optical microscope | Nikon | NIKON ECLIPSE E100 | |
| Urethane | Macklin | 2621-79-6 | |
| Vortex Mixer | Servicebio | MV-100 | |
| Xylene | SCRC | 10023418 |
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